Description
The 145-2C11 monoclonal antibody specifically binds to the 25-kDa ε chain of the T-cell receptor-associated CD3 complex that is expressed on thymocytes, mature T lymphocytes, and NK-T cells. The cytoplasmic domain of CD3e participates in the signal transduction events that activate several cellular biochemical pathways as a result of antigen recognition. Soluble 145-2C11 antibody can activate either unprimed (naive) or primed (memory/preactivated) T cells in vivo or in vitro, in the presence of Fc receptor-bearing accessory cells. In contrast, plate-bound 145-2C11 can activate T cells in the absence of accessory cells. Soluble 145-2C11 antibody has been reported to induce re-directed lysis of Fc receptor-bearing target cells by CTL clones and can also block lysis of specific target cells by antigen-specific CTL's. Under some conditions, T-cell activation by 145-2C11 antibody has been reported to result in apoptotic cell death. The 145-2C11 antibody does not cross-react with rat leukocytes. Preincubation of thymus cell suspensions at 37°C for 2-4 hours prior to staining reportedly enhances the ability of anti-CD3ε and anti-αβ TCR mAbs to detect the T-cell receptor on immature thymocytes.
Immunohistochemistry: The 145-2C11 antibody is recommended to test for immunohistochemical staining of acetone-fixed frozen sections and zinc-fixed paraffin sections. Tissues tested were mouse spleen and thymus. The antibody stains the membranes of all mature T lymphocytes. The isotype control recommended for use with this antibody is purified hamster IgG, group 1, κ (Cat. No. 550344). For optimal indirect immunohistochemical staining, the 145-2C11 antibody should be titrated (1-10 to 1-50 dilution) and visualized via a three-step staining procedure in combination with biotinylated anti-hamster cocktail (Cat. No. 550335) as the secondary antibody and Streptravidin-HRP (Cat. No. 550946) together with the DAB detection system (Cat. No. 550880). The clone 145-2C11 is not recommended for formalin-fixed paraffin embedded sections. For a detailed protocol, please visit the protocols section on our website at http://www.bdbiosciences.com/support/resourse.
Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
1. Although hamster immunoglobulin isotypes have not been well defined, BD Biosciences Pharmingen has grouped Armenian and Syrian hamster IgG monoclonal antibodies according to their reactivity with a panel of mouse anti-hamster IgG mAbs. A table of the hamster IgG groups, Reactivity of Mouse Anti-Hamster Ig mAbs, may be viewed at http://www.bdbiosciences.com/documents/hamster_chart_11x17.pdf.
2. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
4. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
5. This antibody has been developed for the immunohistochemistry application. However, a routine immunohistochemistry test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
6. An isotype control should be used at the same concentration as the antibody of interest.
7. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
